Topical compositions of cyclic amides as immunotherapeutic agents

ABSTRACT

Novel amides and imides are inhibitors of tumor necrosis factors and phosphodiesterase and can be used to combat cachexia, endotoxic shock, retrovirus replication, asthma, and inflammatory conditions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a divisional of Ser. No. 09/007,135 filed Jan. 14, 1998, whichis a continuation of both (1) Ser. No. 08/520,710 filed Aug. 29, 1995,issued as U.S. Pat. No. 5,728,845; and (2) Ser. No. 08/578,738 filedDec. 26, 1995, issued as U.S. Pat. No. 5,728,844, the disclosures ofwhich are incorporated herein by reference. Ser. No. 08/578,738 is acontinuation-in-part of Ser. No. 08/520,710.

BACKGROUND OF THE INVENTION

This invention relates to a method of reducing the level of cytokinesand their precursors in mammals and to compounds and compositions usefultherein.

In particular, the invention pertains to a class of compounds whichinhibit the action of phosphodiesterases, particularly PDE III and PDEIV, and the formation of TNFα and NF_(κ)B. In a first embodiment, thecompounds of the present invention can be diagrammatically representedby the formula:

in which:

R⁵ is:

-   -   (i) the divalent residue of pyridine, pyrrolidine, imidizole, or        thiophene, wherein the two bonds of the divalent residue are on        vicinal ring carbon atoms;    -   (ii) a divalent cycloalkyl of 4 to 10 carbon atoms,        unsubstituted or substituted with one or more stibstituents each        selected independently of the other from the group consisting of        nitro, cyano, trifluoromethyl, carbethoxy, carbomethoxy,        carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy,        amino, substituted amino, alkyl of 1 to 10 carbon atoms, alkoxy        of 1 to 10 carbon atoms, phenyl or halo;    -   (iii) di-substituted vinylene, substituted with nitro, cyano,        trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,        carbamoyl, carbamoyl substituted with and alkyl of 1 to 3 carbon        atoms, acetoxy, carboxy, hydroxy, amino, amino substituted with        an alkyl of 1 to 3 carbon atoms, alkyl of 1 to 4 carbon atoms,        alkoxy of 1 to 4 carbon atoms, or halo; or    -   (iv) ethylene, unsubstituted or substituted with 1 to 2        substituents each selected independently from nitro, cyanb,        trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,        carbanoyl, carbamoyl substituted with and alkyl of 1 to 3 carbon        atoms, acetoxy, carboxy, hydroxy, amino, amino, substituted with        an alkyl of 1 to 3 carbon atoms, alkyl of 1 to 4 carbon atoms,        alkoxy of 1 to 4 carbon atoms, or halo;

R⁶is —CO—, —CH₂—, —CH₂CO—, or —SO₂—;

R⁷ is

-   -   (i) cyclic or bicyclic alkyl of 4 to 12 carbon atoms;    -   (ii) pyridyl;    -   (iii) phenyl substituted with one or more substituents each        selected independently of the other from nitro, cyano,        trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,        carbamoyl, acetoxy, carboxy, hydroxy, amino, straight or        branched alkyl of 1 to 10 carbon atoms, straight or branched        alkoxy of 1 to 10 carbon atoms, or halo;    -   (iv) benzyl substituted with one to three substituents each        selected independently from the group consisting of nitro,        cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy,        acetyl, carbamoyl, acetoxy, carboxy, hydroxy, amino, alkyl of 1        to 4 carbon atoms, alkoxy of 1 to I0 carbon atoms, or halo;    -   (v) naphthyl; or    -   (vi) benzyloxy;

Y is —COX, —C≡N, —OR⁸, alkyl of 1 to 5 carbon atoms, or aryl;

X is —NH₂, —OH, —NHR, —R⁹, —OR⁹, or alkyl of 1 to 5 carbon atoms;

R⁸ is hydrogen or lower alkyl;

R⁹ is alkyl or benzyl; and,

n has a value of 0, 1, 2,or3.

Within this group, Y is preferably —C≡N or —CO(CH₂)_(m) CH₃ in which mhas a value of 0, 1, 2, or 3; and

In a second embodiment, the compounds of the present invention can bediagrammatically represented by the formula:

in which:

one of R¹ and R² is R³—X— and the other is hydrogen, nitro, cyano,trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,carbamoyl, acetoxy, carboxy, hydroxy, amino, lower alkyl, lower alkoxy,halo, or R³—X—;

R3 is monocycloalkyl of up to 10 carbon atoms, polycycloalkyl of up to10 carbon atoms, or benzocyclic alkyl of up to 10 carbon atoms;

X is —CH₂— or —O—;

R⁵ is:

-   -   (i) the vicinally divalent residue of pyridine, pyrrolidine,        imidizole, or thiophene, wherein the two bonds of the divalent        residue are on vicinal ring carbon atoms,    -   (ii) a vicinally divalent cycloalkyl of 4-10 carbon atoms,        unsubstituted or substituted with 1 to 3 substituents each        selected independently from the group consisting of nitro,        cyano, halo, trifluoromethyl, carbethoxy, carbomethoxy,        carbopropoxy, acetyl, carbamoyl, acetoxy, carboxy, hydroxy,        amino, substituted amino, alkyl of 1 to 10 carbon atoms, alkoxy        of 1 to 10 carbon atoms, phenyl;    -   (iii) di-substituted vinylene, substituted with nitro, cyano,        trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,        carbamoyl, carbamoyl substituted with and alkyl of 1 to 3 carbon        atoms, acetoxy, carboxy, hydroxy, amino, amino substituted with        an alkyl of 1 to 3 carbon atoms, alkyl of 1 to 4 carbon atoms,        alkoxy of 1 to 4 carbon atoms, or halo; or    -   (iv) ethylene, unsubstituted or substituted with 1 to 2        substituents each selected independently from nitro, cyano,        trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,        carbamoyl, carbamoyl substituted with and alkyl of 1 to 3 carbon        atoms, acetoxy, carboxy, hydroxy, amino, anino substituted with        an alkyl of 1 to 3 carbon atoms, alkyl of 1 to 4 carbon atoms,        alkoxy of 1 to 4 carbon atoms, or halo;

R⁶ is —CO—, —CH₂—, or —CH₂CO—;

Y is —COX, —C≡N, —OR⁸, alkyl of 1 to 5 carbon atoms, or aryl;

X is —NH₂, —OH, —NHR, —R⁹, —OR⁹, or alkyl of 1 to 5 carbon atoms;

R⁸ is hydrogen or lower alkyl;

R⁹ is alkyl or benzyl; and,

n has a value of 0, 1, 2, or 3.

The term alkyl as used herein denotes a univalent saturated branched orstraight hydrocarbon chain. Unless otherwise stated, such chains cancontain from 1 to 18 carbon atoms. Representative of such alkyl groupsare methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl,tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, hexyl, isohexyl,heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl,pentadecyl, hexadecyl, heptadecyl, octadecyl, and the like. Whenqualified by “lower”, the alkyl group will contain from 1 to 6 carbonatoms. The same carbon content applies to the parent term “alkane” andto derivative terms such as “alkoxy”.

The term cycloalkyl as used herein denotes a univalent saturated cyclichydrocarbon chain. Unless otherwise stated, such chains can contain upto 18 carbon atoms. Monocyclicalkyl refers to groups having a singlering group. Polycycloalkyl denotes hydrocarbon systems containing two ormore ring systems with two or more ring carbon atoms in common.Benzocycloalkyl signifies a monocyclicalkyl group fused to a benzogroup.

Representative of monocycloalkyl groups are cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl,cyclodecyl, cycloundecyl, cyclododecyl, cyclotridecyl, cyclotetradecyl,cyclopentadecyl, cyclohexadecyl, cycloheptadecyl, and cyclooctadecyl.Representative of polycycloalkyl include bicyclo[2.2.1]heptyl,bicyclo[3.2.1]octyl, and bicyclo[2.2.2]octyl. Benzocycloalkyl istypified by tetrahydronaphthyl, indanyl, and 1.2-benzocycloheptanyl.

Tumor necrosis factor α, or TNFα, is a cytokine which is releasedprimarily by mononuclear phagocytes in response to a numberimmunostimulators. When administered to animals or humans, it causesinflammation, fever, cardiovascular effects, hemorrhage, coagulation,and acute phase responses similar to those seen during acute infectionsand shock states. Excessive or unregulated TNFα production thus has beenimplicated in a number of disease conditions. These include endotoxemiaand/or toxic shock syndrome {Tracey et al., Nature 330, 662-664 (1987)and Hinshaw et al., Circ. Shock 30, 279-292 (1990)}; cachexia {Dezube etal., Lancet, 335 (8690), 662 (1990)} and Adult Respiratory DistressSyndrome where TNFα concentration in excess of 12,000 pg/mL have beendetected in pulmonary aspirates from ARDS patients {Millar et al.,Lancet 2 (8665), 712-714 (1989)}. Systemic infusion of recombinant TNFαalso resulted in changes typically seen in ARDS {Ferrai-Baliviera etal., Arch. Surg. 124 (12), 1400-1405 (1989)}.

TNFα appears to be involved in bone resorption diseases, includingarthritis. When activated, leukocytes will produce bone-resorption, anactivity to which the data suggest TNFα contributes. {Bertolini et al.Nature 319, 516-518 (1986) and Johnson et al., Endocrinology 124 (3),1424-1427 (1989).} TNFα also has been shown to stimulate bone resorptionand inhibit bone formation in vitro and in vivo through stimulation ofosteoclast formation and activation combined with inhibition ofosteoblast function. Although TNFα may be involved in many boneresorption diseases, including arthritis, the most compelling link withdisease is the association between production of TNFα by tumor or hosttissues and malignancy associated hypercalcemia {Calci. Tissue Int. (US)46 (Suppl.), S3-10 (1990)}. In Graft versus Host Reaction, increasedserum TNFα levels have been associated with major complication followingacute allogenic bone marrow transplants {Holler et al., Blood, 75 (4),1011-1016 (1990)}.

Cerebral malaria is a lethal hyperacute neurological syndrome associatedwith high blood levels of TNFα and the most severe complicationoccurring in malaria patients. Levels of serum TNFα correlated directlywith the severity of disease and the prognosis in patients with acutemalaria attacks {Grau et al., N. Engl. J. Med. 320 (24), 1586-1591(1989)}.

Macrophage-induced angiogenesis TNFα is known to be mediated by TNFα.Leibovich et al. {Nature, 329, 630-632 (1987)} showed TNFα induces invivo capillary-blood vessel formation in the rat cornea and thedeveloping chick chorioallantoic membranes at very low doses and suggestTNFα is a candidate for inducing angiogenesis in inflammation, woundrepair, and tumor growth. TNFα production also has been associated withcancerous conditions, particularly induced tumors {Ching et al., Brit.J. Cancer, (1955) 72, 339-343, and Koch, Progress in MedicinalChemistry, 22, 166-242 (1985)}.

TNFα also plays a role in the area of chronic pulmonary inflammatorydiseases. The deposition of silica particles leads to silicosis, adisease of progressive respiratory failure caused by a fibroticreaction. Antibody to TNFα completely blocked the silica-induced lungfibrosis in mice {Pignet et al., Nature, 344, 245-247 (1990)}. Highlevels of TNFα production (in the serum and in isolated macrophages)have been demonstrated in animal models of silica and asbestos inducedfibrosis {Bissonnette et al., Inflammation 13 (3), 329-339 (1989)}.Alveolar macrophages from pulmonary sarcoidosis patients have also beenfound to spontaneously release massive quantities of TNFα as comparedwith macrophages from normal donors {Baughman et al., J. Lab. Clin. Med.115 (1), 36-42 (1990)}.

TNFα is also implicated in the inflammatory response which followsreperfusion, called reperfusion injury, and is a major cause of tissuedamage after loss of blood flow {Vedder et al., PNAS 87, 2643-2646(1990)}. TNFα also alters the properties of endothelial cells and hasvarious pro-coagulant activities, such as producing an increase intissue factor pro-coagulant activity and suppression of theanticoagulant protein C pathway as well as down-regulating theexpression of thrombomodulin {Sherry et al., J. Cell Biol. 107,1269-1277 (1988)}. TNFα has pro-inflammatory activities which togetherwith its early production (during the initial stage of an inflammatoryevent) make it a likely mediator of tissue injury in several importantdisorders including but not limited to, myocardial infarction, strokeand circulatory shock. Of specific importance may be TNFα-inducedexpression of adhesion molecules, such as intercellular adhesionmolecule (ICAM) or endothelial leukocyte adhesion molecule (ELAM) onendothelial cells {Munro et al.. Am. J. Path. 135 (1), 121-132 (1989)}.

TNFα blockage with monoclonal anti-TNFα antibodies has been shown to bebeneficial in rheumatoid arthritis {Elliot et al., Int. J. Pharmac. 199517 (2), 141-145}. High levels of TNFα are associated with Crohn'sdisease {von Dullemen et al., Gastroenterology, 1995 109 (1), 129-135}and clinical benefit has been achieved with TNFα antibody treatment.

Moreover, it now is known that TNFα is a potent activator of retrovirusreplication including activation of HIV-1. {Duh et al., Proc. Nat. Acad.Sci. 86, 5974-5978 (1989); Poll et al., Proc. Nat. Acad. Sci. 87,782-785 (1990); Monto et al., Blood 79, 2670 (1990); Clouse et al., J.Immunol. 142, 431-438 (1989); Pollet al., AIDS Res. Hum. Retrovirus,191-197 (1992)}. AIDS results from the infection of T lymphocytes withHuman Immunodeficiency Virus (HIV). At least three types or strains ofHIV have been identified, ie., HIV-1, HIV-2 and HIV-3. As a consequenceof HIV infection, T-cell mediated immunity is impaired and infectedindividuals manifest severe opportunistic infections and/or unusualneoplasms. HIV entry into the T lymphocyte requires T lymphocyteactivation. Other viruses, such as HIV-1, HIV-2 infect T lymphocytesafter T cell activation and such virus protein expression and/orreplication is mediated or maintained by such T cell activation. Once anactivated T lymphocyte is infected with HIV, the T lymphocyte mustcontinue to be maintained in an activated state to permit HIV geneexpression and/or HIV replication. Cytokines, specifically TNFα, areimplicated in activated T-cell mediated HIV protein expression and/orvirus replication by playing a role in maintaining T lymphocyteactivation. Therefore, interference with cytokine activity such as byprevention or inhibition of cytokine production, notably TNFα, in anHIV-infected individual assists in limiting the maintenance of Tlymphocyte caused by HIV infection.

Monocytes, macrophages, and related cells, such as kupffer and glialcells, also have been implicated in maintenance of the HIV infection.These cells, like T cells, are targets for viral replication and thelevel of viral replication is dependent upon the activation state of thecells. {Rosenberg et al., The Immunopathogenesis of HIV Infection,Advances in Immunology, 57 (1989)}. Cytokines, such as TNFα, have beenshown to activate HIV replication in monocytes and/or macrophages {Poliet al., Proc. Natl. Acad. Sci., 87, 782-784 (1990)}, therefore,prevention or inhibition of cytokine production or activity aids inlimiting HIV progression for T cells. Additional studies have identifiedTNFα as a common factor in the activation of HIV in vitro and hasprovided a clear mechanism of action via a nuclear regulatory proteinfound in the cytoplasm of cells (Osborn, et al., PNAS 86 2336-2340).This evidence suggests that a reduction of TNFα synthesis may have anantiviral effect in HIV infections, by reducing the transcription andthus virus production.

AIDS viral replication of latent HIV in T cell and macrophage lines canbe induced by TNFα {Folks et al., PNAS 86, 2365-2368 (1989)}. Amolecular mechanism for the virus inducing activity is suggested byTNFα's ability to activate a gene regulatory protein (NFκB) found in thecytoplasm of cells, which promotes HIV replication through binding to aviral regulatory gene sequence (LTR) {Osborn et al., PNAS 86, 2336-2340(1989)}. TNFα in AIDS associated cachexia is suggested by elevated serumTNFα and high levels of spontaneous TNFα production in peripheral bloodmonocytes from patients {Wright et al., J. Immunol. 141 (1), 99-104(1988)}. TNFα has been implicated in various roles with other viralinfections, such as the cytomegalia virus (CMV), influenza virus,adenovirus, and the herpes family of viruses for similar reasons asthose noted.

The nuclear factor κB (NFκB) is a pleiotropic transcriptional activator(Lenardo, et al., Cell 1989, 58, 227-29). NFκB has been implicated as atranscriptional activator in a variety of disease and inflammatorystates and is thought to regulate cytokine levels including but notlimited to TNFα and also to be an activator of HIV transcription(Dbaibo, et al., J. Biol. Chem. 1993, 17762-66; Duh et al., Proc. Natl.Acad Sci. 1989, 86, 5974-78; Bachelerie et al., Nature 1991, 350,709-12; Boswas et al., J. Acquired Immune Deficiency Syndrome 1993, 6,778-786; Suzuki et al., Biochem. And Biophys. Res. Comm. 1993, 193,277-83; Suzuki et al., Biochem. And Biophys. Res Comm. 1992, 189,1709-15; Suzuki et al., Biochem. Mol. Bio. Int. 1993, 31 (4), 693-700;Shakhov et al., Proc. Natl. Acad. Sci. USA 1990, 171, 35-47; and Staalet al., Proc. Natl. Acad. Sci. USA 1990, 87, 9943-47). Thus, inhibitionof NFκB binding can regulate transcription of cytokine gene(s) andthrough this modulation and other mechanisms be useful in the inhibitionof a multitude of disease states. The compounds described herein caninhibit the action of NFκB in the nucleus and thus are useful in thetreatment of a variety of diseases including but not limited torheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, otherarthritic conditions, septic shock, septis, endotoxic shock, graftversus host disease, wasting, Crohn's disease, ulcerative colitis,multiple sclerosis, systemic lupus erythrematosis, ENL in leprosy, HIV,AIDS, and opportunistic infections in AIDS. TNFα and NFκB levels areinfluenced by a reciprocal feedback loop. As noted above, the compoundsof the present invention affect the levels of both TNFα and NFκB.

Many cellular functions are mediated by levels of adenosine 3′,5′-cyclicmonophosphate (cAMP). Such cellular functions can contribute toinflammatory conditions and diseases including asthma, inflammation, andother conditions (Lowe and Cheng, Drugs of the Future, 17 (9), 799-807,1992). It has been shown that the elevation of cAMP in inflammatoryleukocytes inhibits their activation and the subsequent release ofinflammatory mediators, including TNFα and NFκB. Increased levels ofcAMP also leads to the relaxation of airway smooth muscle.Phosphodiesterases control the level of cAMP through hydrolysis andinhibitors of phosphodiesterases have been shown to increase cAMPlevels.

Decreasing TNFα levels anid/or increasing cAMP levels thus constitutes avaluable therapeutic strategy for the treatment of many inflammatory,infectious, immunological or malignant diseases. These include but arenot restricted to septic shock, sepsis, endotoxic shock, hemodynamicshock and sepsis syndrome, post ischemic reperfusion injury, malaria,mycobacterial infection, meningitis, psoriasis, congestive heartfailure, fibrotic disease, cachexia, graft rejection, cancer, autoimmunedisease, opportunistic infections in AIDS, rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis, other arthritic conditions,Crohn's disease, ulcerative colitis, multiple sclerosis, systemic lupuserythrematosis, ENL in leprosy, radiation damage, and hyperoxic alveolarinjury. Prior efforts directed to the suppression of the effects of TNFαhave ranged from the utilization of steroids such as dexamethasone andprednisolone to the use of both polyclonal and monoclonal antibodies{Beutler et al., Science 234, 470-474 (1985); WO 92/11383}.

The compounds claimed in this patent inhibit the action of NFκB in thenucleus and thus are useful in the treatment of a variety of diseasesincluding but not limited to rheumatoid arthritis, rheumatoidspondylitis, osteoarthritis, other arthritic conditions, septic shock,septis, endotoxic shock, graft versus host disease, wasting, Crohn'sdisease, ulcerative colitis, multiple sclerosis, systemic lupuserythrematosis, ENL in leprosy, HIV, AIDS, and opportunistic infectionsin AIDS.

It is not known at this time, however, how the compounds of the presentinvention regulate the levels of TNFα, NFκB, or both. As noted above,the compounds of the present invention affect the levels of both TNFαand NFκB.

The compounds can be used, under the supervision of qualifiedprofessionals, to inhibit the undesirable effects of TNFα orphosphodiesterase. The compounds can be administered orally, rectally,or parenterally, alone or in combination with other therapeutic agentsincluding antibiotics, steroids, etc., to a mammal in need of treatment.Oral dosage forms include tablets, capsules, dragees, and similarshaped, compressed pharmaceutical forms. Isotonic saline solutionscontaining 20-100 milligrams/milliliter can be used for parenteraladministration which includes intramuscular, intrathecal, intravenousand intra-arterial routes of administration. Rectal administration canbe effected through the use of suppositories formulated fromconventional carriers such as cocoa butter.

Dosage regimens must be titrated to the particular indication, the age,weight, and general physical condition of the patient, and the responsedesired but generally doses will be from about 1 to about 1000milligrams/day as needed in single or multiple daily administration. Ingeneral, an initial treatment regimen can be copied from that known tobe effective in interfering with TNFα activity for other TNFα mediateddisease states by the compounds of the present invention. Treatedindividuals will be regularly checked for T cell numbers and T4/T8ratios and/or measures of viremia such as levels of reversetranscriptase or viral proteins, and/or for progression ofcytokine-mediated disease associated problems such as cachexia or muscledegeneration. If no effect is observed following the normal treatmentregimen, then the amount of cytokine activity interfering agentadministered is increased, e.g., by fifty percent a week.

The compounds of the present invention can also be used topically in thetreatment or prophylaxis of topical disease states mediated orexacerbated by excessive TNFα production, such as viral infections, forexample those caused by the herpes viruses or viral conjunctivitis,psoriasis, other skin disorders and diseases, etc.

The compounds can also be used in the veterinary treatment of mammalsother than humans in need of prevention or inhibition of TNFαproduction. TNFα mediated diseases for treatment, therapeutically orprophylactically, in animals include disease states such as those notedabove, but in particular viral infections. Examples include felineimmunodeficiency virus, equine infectious anaemia virus, caprinearthritis virus, visna virus, and maedi virus, as well as otherlentiviruses.

These compounds possess at least one center of chirality and thus willexist as optical isomers. Both the racemates of these isomers and theindividual isomers themselves, as well as diastereoisomers when thereare two or more chiral centers, are within the scope of the presentinvention. The racemates can be used as such or can be separated intotheir individual isomers mechanically as by chromatography using achiral absorbent. Alternatively, the individual isomers can be preparedin chiral form or separated chemically from a mixture by forming saltswith a chiral acid, such as the individual enantiomers of10-camphorsulfonic acid, camphoric acid, alpha-bromocamphoric acid,methoxyacetic acid, tartaric acid, diacetyltartaric acid, malic acid,pyrrolidone-5-carboxylic acid, and the like, and then freeing one orboth of the resolved bases, optionally repeating the process, so as toobtain either or both isomers substantially free of the other; i.e., ina form having an optical purity of >95%.

Prevention or inhibition of production of TNFα by these compounds can beconveniently assayed using methods known in the art. For example, TNFαInhibition Assays can be performed as follows:

PBMC isolation: PBMC from normal donors were obtained by Ficoll-Hypaquedensity centrifugation. Cells were cultured in RPMI supplemented with10% AB+ serum, 2mM L-glutamine, 100 U/mL penicillin and 100 mg/mLstreptomycin.

PBMC suspensions: Drugs were dissolved in DMSO (Sigma Chemical), furtherdilutions were done in supplemented RPMI. The final DMSO concentrationin the presence or absence of drug in the PBMC suspensions was 0.25 wt%. Drugs were assayed at half-log dilutions starting at 50 mg/mL. Drugswere added to PBMC (10⁶ cells/mL) in 96 wells plates one hour before theaddition of LPS.

Cell stimulation: PBMC (10⁶ cells/mL) in the presence or absence of drugwere stimulated by treatment with 1 mg/mL of LPS from Salmonellaminnesota R595 (List Biological Labs, Campbell, Calif.). Cells were thenincubated at 37° C. for 18-20 hours. Supernatants were then harvestedand assayed immediately for TNFα levels or kept frozen at −70° C. (fornot more than 4 days) until assayed.

TNFα Determination: The concentration of TNFα in the supernatant wasdetermined by human TNFα ELISA kits (ENDOGEN, Boston, Mass.) accordingto the manufacturer's directions.

Another assay procedure utilizes plates (Nunc Immunoplates, Roskilde,DK) which are treated with 5 mg/mL of purified rabbit anti-TNFαantibodies at 4° C. for 12 to 14 hours. The plates then are blocked for2 hours at 25° C. with PBS/0.05% Tween containing 5milligrams/milliliter BSA. After washing, 100 mL of unknowns as well ascontrols are applied and the plates incubated at 4° C. for 12 to 14hours. The plates are washed and assayed with a conjugate of peroxidase(horseradish) and mouse anti-TNFα monoclonal antibodies, and the colordeveloped with o-phenylenediamine in phosphate-citrate buffer containing0.012% hydrogen peroxide and read at 492 nm.

The compounds can be prepared using methods which are known per se. forexanple, a cyclic anhydride of lactone can be reacted with theappropriate amine:

in which R⁵, R⁶, R^(7,) Y, and n are as defined above. The reaction canbe effected analogously to the methods described in U.K. PatentSpecification No. 1,036,694, the disclosure of which is incorporatedherein by reference. Optionally acetic acid, with or without sodiumacetate, can be added.

In place of the acid anhydride or lactone, one can utilize anN-carbethoxy derivative of the formula:

In a further embodiment, compounds in which R⁶ is —CH₂— can be formedthrough condensation of a dialdehyde with a disubstituted aromaticcompound in the presence of refluxing acetic acid utilizing the methodof Griggs et al., J. Chem. Soc. Chem. Comm., 1985, 1183-1184, thedisclosure of which is incorporated herein by reference.

The disubstituted aromatic starting materials can be obtained throughcondensation of an appropriately substituted aldehyde and malonic acid,with intermediate formation of the amidine and subsequentdecarboxylation.

The disubstituted aldehydes can be prepared utilizing classical methodsfor ether formation; e.g., reaction with the appropriate bromide in thepresence of potassium carbonate. Numerous cycloalkyloxy benzaldehydesand procedures for preparing them are described in the literature. See,e.g., Ashton et al., J. Med. Chem., 1994, 37, 1696-1703; Saccomano etal., J. Med. Chem., 1994, 34,, 291-298; and Cheng et al., Org. and Med.Chem. Lett., 1995, 5 (17), 1969-1972, the disclosures of which areincorporated herein by reference.

Typical compounds include3-(1-oxobenzo[f]isoindol-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid,3-(1-oxobenzo[f]isoindol-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionicacid, 3-(1-oxobenzo[f]isoindol-2-yl)-3-(3-methoxy-4-ethoxyphenyl)propionic acid,3-(1-oxobenzo[f]isoindol-2-yl)-3-(3,4-dimethoxyphenyl)propionic acid,3-(1-oxo-benzo[f]isoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1-oxo-4-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1-oxo-5-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1-oxo-4-azaisoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionicacid, 3-(1-oxo-4-azaisoindol-2-yl)-3-(3-methoxy-4-ethoxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-cyclopentyloxy4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-methoxy-4-cyclobutyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-methoxy-4-cyclbpentyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxo-5-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionamide,3(1,3-di-oxo-4-azaisoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-methoxy-4-cyclobutyloxyphenyl)propionamide,3-(1-oxobenzo[f]isoindol-2-yl)-3-(3-cyclopentyloxy-4-cyclohexyloxyphenyl)propionicacid,3-(1-oxobenzo[f]isoindol-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1-oxobenzo[f]isoindol-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionamide,3-(1-oxobenzo[f]isoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyl)propionamide,3-(1-oxobenzo[f]isoindol-2-yl)-3-(3,4-dicyclopentyloxyphenyl)propionamide,3-(1-oxobenzo[f]isoindol-2-yl)-3-(3,4-dicyclohexyloxyphenyl)propionamide,3-(1-oxo-4-azalsoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionamide,3-(1-oxo-5-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionamide,3-(1-oxo-4-azaisoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionamide,3-(1-oxo-4-azaisoindol-2-yl)-3-(3-cyclobexyloxy-4-ethoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionic acid,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionicacid, 3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionic acid,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3,4-dicyclohexyloxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3,4-dicyclopentyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1,3-dioxo-5-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionicacid, methyl 3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate, methyl3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionate,methyl3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionate,methyl3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3,4-di-cyclopentyloxyphenyl)propionate,methyl3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3,4-dicyclohexyloxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionate,methyl3-(1,3-dioxo-5-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionate,ethyl3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate,ethyl3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionate,ethyl3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3-methoxy-4-ethoxyphenyl)propionate,ethyl3-(1,3-dioxobenzo[f]isoindol-2-yl)-3-(3,4-dimethoxyphenyl)propionate,ethyl3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionate,ethyl3-(1,3-dioxo-5-azaisoindol-2-yl)-3-(3-methoxy-4-cyclopentyloxyphenyl)propionate,ethyl and3-(1,3-dioxo-4-azaisoindol-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionate.

Representative aldehyde starting materials include3-cyclopentyloxy4-methoxybenzaldehyde,3-cyclopentyloxy-4-ethoxybenzaldehyde,3-cyclohexyloxy-4-cyclohexyloxybenzaldehyde,3-(exo-bicyclo[2.2.1]hept2-yloxy)-4-methoxybenzaldehyde,3-(endo-bicyclo[2.2.1]hept-2-yl-oxy)-4-methoxybenzaldehyde,3-(bicyclo[2.2.2]oct-2-yloxy)-4-methoxybenzaldehyde,3-(bi-cyclo[3.2.1]oct-2-yloxy)-4-methoxybenzaldehyde,3-indan-2-yloxy-4-methoxybenzaldehyde, and3-(endo-benzobicyclo[2.2.1]hept-2-yloxy)-4-methoxybenzaldehyde.

The following examples will serve to further typify the nature of thisinvention but should not be construed as a limitation in the scopethereof, which scope is defined solely by the appended claims.

EXAMPLE 1 3-Amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionic Acid

A stirred suspension of 3-cyclopentyloxy-4-methoxybenzaldehyde (10.0 g,45.4 mmol) and ammonium acetate (7.00 g, 90.8 mmol) in ethanol (95%, 30mL) under nitrogen was heated to 45-50° C. and malonic acid (4.72 g,45.4 mmol) was added. The solution was heated at reflux for 24 hours. Awhite solid precipitated, the mixture was allowed to cool to roomtemperature and was then filtered. The white solid was washed withethanol, air dried and then dried in vacuo (60° C., <1 mm) to afford7.36 g (58%) of the product: mp 225-226° C.; ¹H NMR (D₂O/NaOH/TSP)δ7.05-6.88 (m, 3H), 4.91-4.78 (m, 1H), 4.21-4.14 (m, 1H), 3.79 (s, 3H),2.59-2.46 (m, 2H), 2.05-1.48 (m, 8H). Trace impurity peaks were presentat 6.39 and 7.34 ppm. ¹³C NMR (D₂O/NaOD/TSP) δ182.9, 150.7, 149.1,140.6, 121.6, 116.0, 114.9, 83.9, 58.5, 55.3, 49.8, 34.9, 26.3.

Similarly from equivalent amounts of3-ethoxy-4-cyclopentyloxybenzaldehyde,3-ethoxy-4-cyclohexyloxybenzaldehyde,3-methoxy-4-cyclopentyloxybenzaldehyde,3-methoxy-4-cyclohexyloxybenzaldehyde,3-cyclohexyloxy-4-methoxybenzaldehyde,3-cyclopentyloxy-4-ethoxybenzaldehyde, and3-cyclohexyloxy-4-ethoxybenzaldehyde, there are respectively obtainedaccording to the foregoin procedure3-amino-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionic acid,3-amino-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionic acid,3-amino-3-(3-methoxy-4-cyclohexyloxyphenyl)propionic acid,3-amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionic acid,3-amino-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionic acid,3-amino-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionic acid, and3-amino-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionic acid.

EXAMPLE 2 3-Phthalimido-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicAcid

To a stirred mixture of3-amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionic acid (2.34 g. 8.40mmol) and sodium carbonate (0.96 g, 9.05 mmol) in a mixture of water (20mL) and acetonitrile (20 mL) under nitrogen was addedN-carbethoxyphthalimide (1.9 g, 8.4 mmol). After 3 hours, theacetonitrile was removed in vacuo. The pH of the solution was adjustedto 1 with aqueous hydrogen chloride (4 N). Ether (5 mL) was added andthe mixture stirred for 1 hour. The resulting slurry was filtered andthe solid washed with water, air dried and then dried in vacuo (60° C.,<1 mm) to afford 2.92 g (85%) of the product as a white solid: mp159-162° C.; ¹H NMR (DMSO-d₆) δ12.40 (br s, 1H), 7.96-7.80 (m, 4H), 7.02(s, 1H), 6.90 (s, 2H), 5.71-5.52 (m, 1H), 4.81-4.65 (m, 1H), 3.70 (s,3H), 3.59-3.16 (m, 2H), 2.00-1.44 (m, 8H); ¹³C NMR (DMSO-d₆) δ171.7,167.6, 149.1, 146.8, 134.6, 131.2, 131.1, 123.1, 119.4, 113.9, 112.1,79.5, 55.5, 50.1, 36.1, 32.1, 32.1, 23.5; Anal. Calcd for C₂₃H₂₃NO₆.Theoretical: C, 67.47; H, 5.66; N, 3.42. Found: C, 67.34; H, 5.59; N,3.14.

By substituting an equivalent amount of2-carbethoxy-1,3-dioxobenzo[f]isoindoline in the foregoing procedure,there is obtained3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid. Likewise from 2-carbethoxy-1,3-dioxo-4-azaisoindoline and2-carbethoxy-1,3-dioxo-5-azaisoindoline there are respectively obtained3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid and3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid.

1,3-Dioxobenzo[f]isoindoline, obtained from naphthalene-2,3-dicarboxylicacid anhydride through treatment with ammonia, is acylated as with ethylchloroformate to yield 2-carbethoxy-1,3-dioxobenzo[f]isoindoline.Pyridine-2,3-dicarboxylic acid anhydride and pyridine-3,4-dicarboxylicacid anhydride are similarly converted to2-carbethoxy-1,3-dioxo-4-azaisoindoline and2-carbethoxy-1,3-dioxo-5-azaisoindoline.

Use in the procedure of this example of equivalent amounts of2-carbethoxy-1,3-dioxobenzo[f]isoindoline with3-amino-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionic acid,3-amino-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionic acid,3-amino-3-(3-methoxy-4-cyclohexyloxyphenyl)propionic acid,3-amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionic acid,3-amino-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionic acid,3-amino-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionic acid, and3-amino-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionic acid, all preparedas described in Example 1, yield3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid.3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionicacid, and3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionicacid, respectively.

Likewise by employing 2-carbethoxy-1,3-dioxo-4-azaisoindoline there areobtained from the same amines of Example 1 the compounds3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3ethoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-methoxy4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy4-methoxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)4-methoxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionicacid, and3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionicacid.

Similarly from 2-carbethoxy-1,3-dioxo-5-azaisoindoline there areobtained with the amines of Example 1 the compounds3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionicacid, and 3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionic acid, respectively.

EXAMPLE 3 3-Phthalimido-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide

A mixture of 3-phthalimido-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid (2.05 g, 5.00 mmol), 1,1′-carbonyldiimidazole (0.91 g, 5.5 mmol)and 4-dimethylaminopyridine (trace) in THF (20 mL) was stirred for 1.5hours under nitrogen at approximately 25° C. To the solution was addedammonium hydroxide (1.07 mL, 16.0 mmol, 28-30%) and stirring wascontinued for 1.5 hours. A small amount of solid precipitated duringthis time. The mixture was concentrated to half its volume and a whitesolid precipitated. The mixture was filtered, washed with a small amountof THF, air dried and dried in vacuo (60° C., <1 mm) to afford 1.27 g ofthe crude product. The crude product was purified by flash columnchromatography (silica gel, 5% MeOH/CH₂Cl₂) and the resulting whitesolid was dried in vacuo (60° C., <1 mm) to afford 1 g (49%) of theproduct: mp 165-166° C.; ¹H NMR (CDCl₃) δ7.85-7.61 (m, 4H), 7.16-7.04(m, 2H), 6.85-6.75 (m, 1H), 5.80 (dd, J=5.8, 10.4 Hz, 1H), 5.66 (br s,1H), 5.54 (br s, 1H), 4.82-4.70 (m, 1H), 3.80 (s, 3H), 3.71 (dd, J=10.4,15 Hz, 1H), 3.06 (dd, J=5.8, 15 Hz, 1H), 2.06-1.51 (m, 8H); ¹³ C NMR(CDCl₃) δ171.8, 168.3, 149.8, 147.7, 133.9, 131.8, 131.3, 123.3, 119.9,114.6, 111.8, 80.4, 56.0, 51.6, 37.9, 32.7, 24.1; Anal. Calcd forC₂₃H₂₄N₂O₅. Theoretical: C, 67.63; H, 5.92; N, 6.86. Found: C, 67.25; H,5.76; N, 6.68.

Similarly from equivalent amounts of3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionic acid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionicacid,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionicacid,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxypheny()propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionicacid,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentylexy-4-ethoxyphenyl)propionicacid, and3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionicacid, there are respectively obtained according to the forgoingprocedure3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide.3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-methoxy4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy4-methoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)4-methoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)4-ethoxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionamide,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionamide, and3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionamide.

EXAMPLE 4 Methyl 3-amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionateHydrochloride

To a cooled (ice bath temperatures) and stirred mixture of3-amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionic acid (3.00 g, 10.7mmol) in methanol (20 mL) under nitrogen was added thionyl chloride (1.8mL, 2.3 mmol) dropwise via syringe. The resulting solution was stirredat 0° C. for 1 hour, the ice bath was removed and stirring was continuedat room temperature for 1 hour and a white solid precipitated. Themethanol was removed and the solid was slurred in hexane. The mixturewas filtered and the white solid was washed with hexane, air dried andthen dried in vacuo (60° C., <1 mm) to afford 2.69 g (76%) of theproduct: mp 183-184.5° C.; ¹H NMR (DMSO-d₆) δ8.76 (br s, 3H), 7.25 (s,1H), 7.06-6.89 (m, 2H), 4.85-4.75 (m, 1H), 4.58-4.44 (m, 1H), 3.74 (s,3H), 3.55 (s, 3H), 3.31-2.86 (m, 2H), 2.06-1.44 (m, 8H); ¹³C NMR(DMSO-d₆) δ169.1, 149.3, 146.5, 128.4, 119.5, 113.5, 111.4, 79.0, 55.0,51.2, 50.3, 38.2, 31.7, 31.6, 23.0; Anal. Calcd for C₁₆H₂₄ClNO₄.Theoretical: C, 58.27; H, 7.33; N. 4.25. Found: C, 58.44; H, 7.34; N,4.13.

Similarly prepared from3-amino-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionic acid,3-amino-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionic acid,3-amino-3-(3-methoxy-4-cyclohexyloxyphenyl)propionic acid,3-amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionic acid,3-amino-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionic acid,3-amino-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionic acid, and3-amino-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionic acid are methyl3-amino-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionate, methyl3-amino-3-(3-ethoxy-4-cyclobexyloxyphenyl)propionate, methyl3-amino-3-(3-methoxy-4-cyclohexyloxyphenyl)propionate, methyl3-amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate, methyl3-amino-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionate, methyl3-amino-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionate, and methyl3-amino-3-(3-cyclohexyloxy)4-ethoxyphenyl)propionate.

EXAMPLE 5 Methyl3-phthalimido-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate

To a stirred solution of methyl3-amino-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate hydrochloride(0.50 g, 1.52 mmol) and sodium carbonate (0.16 g, 1.52 mmol) in amixture of water (5 mL) and acetonitrile (5 mL) under nitrogen was addedN-carbethoxyphthalimide (0.34 g, 1.52 mmol). The solution was stirredfor 3 hours at RT. The acetonitrile was removed in vacuo which affordeda two layer mixture which was extracted with CH₂Cl₂ (3×15 mL). Thecombined organic extracts were dried over MgSO₄, filtered and thenconcentrated in vacuo to afford 0.77 g of the crude product as an oil.The crude product was purified by flash column chromatography (silicagel, 35/65, ethyl acetate/hexane) the resulting glassy solid was driedin vacuo to afford 0.48 g (75%) of the product as a white solid: mp76-78° C.; ¹H NMR (CDCl₃) δ7.86-7.60 (m, 4H), 7.19-7.00 (m, 2H),6.88-6.72 (m, 1H), 5.84-5.67 (m, 1H), 4.85-4.70 (m, 1H), 3.80 (s, 3H),3.80-3.69 (m, 1H), 3.63 (s, 3H), 3.34-3.15 (m, 1H), 2.10-1.48 (m, 8H);¹³C NMR (CDCl₃) δ171.0, 168.0, 149.8, 147.6, 133.9, 131.8, 130.9, 123.2,120.1, 114.6, 111.7, 80.4, 55.9, 51.8, 50.7, 35.9, 32.7. 24.0; Anal:Calcd for C₂₄H₂₅NO₆. Theoretical: C, 68.03; H, 5.95; N, 3.31. Found: C,67.77; H, 5.97; N, 3.20.

Similarly from equivalent amounts of2-carbethoxy-1,3-dioxobenzo[f]isoindoline,2-carbethoxy-1,3-dioxo-4-azaisoindoline, and2-carbethoxy-1,3-dioxo-5-azaisoindoline for N-carbethoxyphthalimide,there are respectively obtained according to the forgoing proceduremethyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate,and methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate.

Use in the procedure of this example of equivalent amounts of2-carbethoxy-1,3-dioxobenzo[f]isoindoline with methyl3-amino-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionate, methyl3-amino-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionate, methyl3-amino-3-(3-methoxy-4-cyclohexyloxyphenyl)propionate, methyl3-amino-3-(3-cyclopentyloxy4-methoxyphenyl)propionate, methyl3-amino-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionate, methyl3-amino-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionate, and methyl3-amino-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionate yields methyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionate,methyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy4-cyclohexyloxyphenyl)propionate,methyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionate,methyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate,methyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionate,methyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionate,and methyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionate.

Likewise by employing 2-carbethoxy-1,3-dioxo-4-azaisoindoline there areobtained from the same amines of Example 4 the compounds methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionate,and methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionate.

Similarly from 2-carbethoxy-1,3-dioxo-5-azaisoindoline there areobtained with the amines of Example 4 the compounds methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionate,methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionate,methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionate,methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionate,methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionate,methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionate,and methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionate.

EXAMPLE 63-Amino-3-(3-{exobicyclo[2.2.1]hept-2-yloxy}-4-methoxyphenyl)propionicAcid

A stirred suspension of3-(exobicyclo[2.2.1]hept-2-yloxy)-4-methoxybenzaldehyde (6.00 g, 24.4mmol) and ammonium acetate (3.76 g, 48.8 mmol) in ethanol (95%, 20 mL)under nitrogen was heated to 45-50° C. and malonic acid (2.53 g, 24.4mmol) was added. The solution was refluxed for 24 hours, allowed to coolto room temperature, and filtered. The solid was washed with ethanol,air dried, and dried in vacuo (60° C., <1 mm) to afford 3.17 g (43%) ofthe product: mp 225-226° C.; ¹H NMR (D₂O/NaOD/TSP) δ7.09-6.90 (m, 3H),4.41-4.28 (m, 1H), 4.27-4.15 (m, 1H), 3.82 (s, 3H), 2.64-2.48 (m, 2H)2.44 (s, 1H), 2.31 (s, 1H), 1.92-1.76 (m, 1H), 1.69-1.38 (m. 4H),1.30-1.05 (m, 3H).

EXAMPLE 7 Methyl3-Amino-3-(3-{exobicyclo[2.2.1]hept-2-yloxy}-4-methoxyphenyl)propionateHydrochloride

To an ice bath cooled stirred suspension of3-amino-3-(3-{exobicyclo[2.2.1]hept-2-yloxy}-4-methoxyphenyl)propionicacid (2.00 g, 6.55 mmol) in methanol (15 mL) under nitrogen was addedthionyl chloride (1.56 mL, 13.l mmol) dropwise via syringe. Theresulting solution was stirred at 0° C. for 30 minutes, the ice bath wasremoved and stirring was continued at room temperature for 2.5 hours.The methanol was removed and the solid slurred in hexane (15 mL). Themixture was filtered and the white solid washed with hexane, air driedand then dried in vacuo (60° C., <1 mm) to afford 1.97 g (85%) of theproduct: mp 197.5-201.5° C.; ¹H NMR (DMSO-d₆) δ7.50 (br s, 3H), 7.18 (s,1H), 7.07-6.88 (m, 2H), 4.56-4.42 (m, 1H), 4.30-4.19 (m, 1H), 3.74 (s,3H), 3.54 (s, 3H), 3.41-2.85 (m, 3H), 2.37 (s, 1H), 2.27 (s, 1H),1.92-1.75 (m, 1H), 1.64-1.03 (m, 6H); ¹³C NMR (DMSO-d₆) δ169.4, 149.6,146.4, 128.8, 120.0, 119.9, 113.8, 111.8, 80.1, 79.9, 55.5, 51.6, 50.7,40.5, 39.2, 38.6, 34.8, 27.8, 23.7, 23.6.

EXAMPLE 8 Methyl3-Phthalimido-3-(3-{exobicyclo[2.2.1]hept-2-yloxy}-4-methoxyphenyl)propionate

To a stirred solution of methyl3-amino-3-(3-{exobicyclo[2.2.1]hept-2-yloxy}-4-methoxyphenyl)propionatehydrochloride (1.00 g, 2.81 mmol) and sodium carbonate (0.3 g, 2.8 mmol)in a mixture of water (10 mL) and acetonitrile (10 mL) under nitrogenwas added N-carbethoxyphthalimide (0.64 g, 2.81 mmol). The solution wasstirred for 3 hours at room temperature. The acetonitrile was remove invacuo and the residue extracted with methylene chloride (3×30 ml). Thecombined organic extracts were dried over MgSO₄, filtered andconcentrated in vacuo to afford 1.44 g of the product. The product wasfurthier purified by flash column chromatography (silica gel, 20%, ethylacetate/methylene chloride) to afford a white solid which was then driedin vacuo to afford 0.23 g (18%) of product: mp 47-48° C.; ¹H NMR (CDCl₃)δ7.86-7.61 (m, 4H), 7.14-7.00 ( m, 2H), 6.82-6.74 (m, 1H), 5.75 (dd,J=5.9, 10 Hz, 1H), 4.25-4.14 (m, 1H), 3.84-3.69 (m, 1H), 3.79 (s, 3H),3.63 (s, 3H), 3.23 (dd, J=5.9, 16.5 Hz, 1H), 2.51-2.41 (m, 1H),2.34-2.24 (m, 1H), 1.86-1.06 (m, 8H); ¹³C NMR (CDCl₃) δ171.1, 168.1,149.7, 147.2, 133.9, 131.8, 130.9, 123.3, 120.1, 120.0, 114.5, 114.4,111.8, 81.1, 56.0, 51.9, 50.8, 41.1, 41.0, 39.9, 39.8, 35.9, 35.5, 35.3,28.4, 24.3; HPLC 97%; Anal. Calcd for C₂₆H₂₇NO₆. Theoretical: C, 69.47;H, 6.05; N, 3.12. Found: C, 69.22; H, 5.91; N, 2.95.

Similarly from equivalent amounts of2-carbethoxy-1,3-dioxobenzo[f]isoindoline,2-carbethoxy-1,3-dioxo-4-azaisoindoline, and2-carbethoxy-1,3-dioxo-5-azaisoindoline for N-carbethoxyphthalimide,there are respectively obtained according to the forgoing proceduremethyl3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-{exobicyclo[2.2.1]hept-2-yloxy}-4-methoxyphenyl)propionate,methyl3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-{exobicyclo[2.2.1]hept-2-yloxy}-4-methoxyphenyl)propionate,and methyl3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-{exobicyclo[2.2.1]hept-2-yloxy}-4-methoxyphenyl)propionamide.

EXAMPLE 9 3-Phthalimido-3-(3,4-diethoxyphenyl)propionitrile

To an ice bath cooled stirred suspension of3-phthalimido-3-(3,4diethoxyphenyl)propionamide (0.96 g, 2.5 mmol),prepared for example as described in U.S. Pat. No. 5,463,063, and4-methylmorpholine (0.66 mL, 6 mmol) in dimethylformamide (9 mL) undernitrogen, was added thionyl chloride (0.35 mL, 4.8 nimol) dropwise.There to was a slight exotherm after which the mixture was stirred at0-5° C. for 30 minutes and at room temperature for 2 hours. The reactionwas monitored by HPLC (Waters Nova-Pak/C-18 column, 3.9×150 mm, 4micron, 1 mL/min, 240 nm, 50/50 acetonitrile/phosphoric acid 0.1% (aq)).The reaction mixture was poured into a mixture of sodium bicarbonate(8.5 mL) and ice (40 g). and stirred until the ice had melted. Themixture was filtered and the solid was washed with copious amounts ofwater. The wet solid was dissolved in methylene chloride (25 mL) and theorganic layer was separated and dried over MgSO₄ and concentrated invacuo to a sticky semi-solid. The solid was purified twice by flashcolumn chromatography (silica gel, 3% ethyl acetate/methylene chloride)to afford a solid which was dried in vacuo (50° C., <1 mm) to afford 0.5g (55 %) of product as a pale yellow solid; ¹H NMR (CDCl₃) δ7.91-7.65(m, 4H), 7.12-6.98 (m, 2H), 6.90-6.78 (m, 1H), 5.61 (dd, J=6.4, 10.3 Hz,1H), 4.19-3.96 (m, 4H),3.83 (dd, J=10.3, 16.8 Hz, 1H), 3.26 (dd, J=6.4,16.8 Hz, 1H), 1.55-1.30 (m, 6H); ¹³C NMR (CDCl₃) δ167.7, 149.2, 148.9,134.3, 131.5, 129.1, 123.6, 120.2. 116.9, 113.2, 112.9, 64.7, 64.5,51.1, 21.1, 14.7; HPLC 98.4 %. Anal. Calcd for C₂₁H₂₀N₂O₄: Theoretical:C, 69.22; H, 5.53; N, 7.69. Found: C, 69.06; H, 5.48; N, 7.58.

Similarly obtained from 3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxotenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionamide,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionamide,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionamide,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionamide,and3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionamideare3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionitrile,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionitrile,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionitrile,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionitrile,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionitrile,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-methoxy4-cyclohexyloxyphenyl)propionitrile,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionitrile,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionitrile,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionitrile,3-(1,3-dioxobenzo[f]isoindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionitrile,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionitrile,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionitrile,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionitrile,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionitrile,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionitrile,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionitrile,3-(1,3-dioxo-4-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionitrile,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclopentyloxyphenyl)propionitrile,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-ethoxy-4-cyclohexyloxyphenyl)propionitrile,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-methoxy-4-cyclohexyloxyphenyl)propionitrile,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-methoxyphenyl)propionitrile,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-methoxyphenyl)propionitrile,3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclopentyloxy-4-ethoxyphenyl)propionitrile,and3-(1,3-dioxo-5-azaindolin-2-yl)-3-(3-cyclohexyloxy)-4-ethoxyphenyl)propionitrile.

EXAMPLE 10 3-Phthalimido-3-(3,4-dimethoxyphenyl)propionitrile

To an ice bath cooled stirred suspension of3-phthalimido-3-(3,4-dimethoxyphenyl)propionamide (1.77 g, 5.00 mmol)and 4-methylmorpholine (1.3 mL, 12 mmol) in dimethyformamide (17 mL)under nitrogen, was added thionyl chloride (0.7 mL, 9.6 mmol) dropwisevia a syringe. There was a slight exotherm and after 30 minutes, thecooling bath was removed and the reaction nlixture was stirred for 2hours at room temperature. The reaction mixture was poured into amixture of sodium bicarbonate (17 g) and 75 mL of ice water and stirreduntil the ice had melted. The slurry was filtered and the solid waswashed with copious amounts of water. The wet solid was dissolved inmethylene chloride (50 mL) and the organic layer was separated, driedover sodium sulfate, and concentrated in vacuo to afford an orangesolid. The solid was purified by flash column chromatography (silicagel, 5/95 ethyl acetate/methylene chloride, 50 mm id column) to afford1.32 g (79%) of the product as a white solid: ¹H NMR (CDCl₃) δ7.9-7.6(m, 4H), 7.10 (m, 2H), 6.83 (m, 1 H), 5.64 (dd, J=6.5, 10.2 Hz, 1H),3.88 (s, 3H), 3.85 (s, 3H), 3.82 (dd, 1H), 3.30 (dd, J=6.5, 16.8 Hz, 1H); ¹³C NMR (CDCl₃) δ167.7, 149.5, 149.2, 134.4, 131.5, 129.1, 123.6,120.1, 116.9, 111.1, 110.7, 56.0, 55.9, 51.1, 21.1. Anal. Calcd forC₁₉H₁₆N₂O₄ 0.18 H₂O. Theoretical: C, 76.2. H, 4.85; N, 8.25. Found: C,67.23; H, 4.79; N, 8.27.

EXAMPLE 11

A stirred mixture 3-amino-3-phenylpropionic acid andcis-1,2-cyclohexanedicarboxylic anhydride in 10 mL of acetic acid undernitrogen was heated to reflux for 4 h and then allowed to cool to roomtemperature. The resulting mixture was concentrated to an orange yellowoil. This oil was crystallized from a 1/1 mixture of ethylacetate/hexane to afford 1.77 g (58%) of3-(cis-hexahydrophthalimido)-3-phenylpropionic acid as white crystals:¹H NMR (DMSO-d₆) δ12.45 (br s, 1 H, COOH), 7.33 (m, 5 H, Ph), 5.48 (dd,1 H, J=6.3, 9.6, CH), 3.41 (dd, 1 H, J=16.5, 9.6 Hz), 3.14 (dd, 1H,J=16.5, 6.3 Hz), 2.50 (m, 2 H), 1.8-1.1 (m, 8 H); ¹³C NMR (DMSO-d₆)δ179.3, 179.2, 171.7, 138.7, 128.4, 127.5, 126.8, 50.1, 38.7, 38.6,35.2. 23.0, 22.9, 21.1. Anal. Calcd for C₁₇H₁₉NO₄. Theory: C, 67.76; H,6.36; N, 4.65. Found: C, 67.52; H, 6.20; N, 4.60.

EXAMPLE 12

A mixture of 3-(cis-hexahydrophthalimido)-3-phenylpropionic acid (0.903g, 3.00 mmol) and carbonyldiimidazole (0.525 g, 3.75 mmol) in 13 mL ofanhydrous tetrahydrofuran under nitrogen was stirred for 1 hour, then0.25 mL of concentrated ammonium hydroxide was added to the reactionsolution. After 20 minutes, the reaction mixture was concentrated invacuo to an oil. The oil was diluted with 20 mL of water and the mixtureextracted with ethyl acetate (20 mL). The organic layer was dried(sodium sulfate) and concentrated to afford an oil. The oil was thenpurified by flash chromatography (silica gel, 5/95 methanol/methylenechloride, R_(f)0.3) to afford 210 mg of3-(cis-hexahydrophthalimido)-3-phenylpropionamide as an oil which slowlycrystallized to an ivory solid: ¹H NMR (DMSO-d₆) d7.49 (s. 1 H, NH),7.4-7.2 (m, 5 H, Ar). 6.90 (s, 1 H, NH), 5.54 (t, 1 H, J 7.8 Hz, CH),3.09 (d, 2 H, J=7.8 Hz, CH2), 2.95-2.80 (m, 2 H, CH2), 1.8-1.1 (m, 8 H);¹³C NMR (DMSO-d₆) δ179.6, 179.5, 171.5, 139.5, 128.6, 127.7, 127.2,55.2, 50.6, 38.8, 36.5, 23.4, 23.3, 21.5

EXAMPLE 13

A stirred mixture of cis-5-norbonene-endo-2,3-dicarboxylic anhydride(1.64 g, 10.0 mmol) and 3-amino-3-phenylpropionic acid (1.65 g, 10.0mmol) in 15 mL of acetic acid under nitrogen was heated to reflux for 6hours. The resulting reaction solution was concentrated in vacuo to anoil which was crystallized from a 1/1 mixture of ethyl acetate/hexane toafford 2.03 g (65%) of 3-(cis-5-norbonene-endo-2,3-dicarboxylicimide)-3-phenylpropionic acid as a white powder: ¹H NMR (DMSO-d₆) δ12.41(br s, 1 H, COOH), 7.29 (m, 5 H, Ph), 6.0-5.7 (m, 2 H), 5.37 (t, 1 H,J=7.7 Hz), 3.5-3.1 (m, 6 H), 1.49 (m, 2 H); ¹³CNMR (DMSO-d₆) δ177.2,177.1, 171.4, 138.3, 134.3, 134.0, 128.1, 127.5, 127.1, 51.4, 50.1,44.8, 44.5, 44.4, 35.1. Anal. Calcd for C₈H₁₇NO₄. Theory: C, 69.44; H,5.50; N, 4.50. Found: C, 69.10; H, 5.33; N,4.43.

EXAMPLE 14

A stirred suspension of 3-amino-3-(3,4-dimethoxyphenyl)propionic acidhydrochloride (0.689 g, 2.50 mmol) and 4-pyridyldicarboxylic acidanhydride (0.373 g, 2.50 mmol) in 20 mL of acetic acid was refluxed forovernight. The cooled reaction was filtered to remove a trace amount ofsolid and the filtrate concentrated to a thick yellow oil. The oil wasdiluted with 20 mL of ethyl acetate and heated to reflux and allowed tocool to room temperature. The resulting slurry was filtered and thefiltrate concentrated to afford a yellow oil which was purified by flashchromatography (silica gel, 2/8 ethyl acetate/methylene chloride) toafford 0.592 g (64%) of methyl3-(1,3-dioxo-5-azaisoindol-2-yl)-3-(3,4-dimethoxyphenyl)-propionate as ayellow oil which slowly solidified to afford a very pale yellow solid:¹H NMR (DMSO-d₆) δ8.15-7.75 (m, 8 H, Ar), 7.75-7.4 (m, 4 h, Ar andCONH), 9.13 (s, 1 H, Ar), 9.11 (d, 1 H, J=4.8 Hz), 7.90 (d, 1 H, J=4.8Hz), 7.03 (s, 1 H), 6.93 (m, 2 H), 5.67 (overlapping dd, 1 H), 3.74 (s,3 H), 3.73 (s, 3 H). 3.56 (s, 3 H), 3.65-3.30 (m, 2 H); ¹³C NMR(DMSO-d₆) δ170.7, 166.9, 166.5, 156.0, 148.6, 148.5, 144.1, 138.7,130.4, 125.2, 119.1, 116.9, 111.6, 111.1, 55.4, 51.6, 50.1, 35.4.

EXAMPLE 15

Tablets, each containing 50 milligrams of active ingredient, can beprepared in the following manner:

Constituents (for 1000 tablets)

active ingredient 50.0 grams lactose 50.7 grams wheat starch 7.5 gramspolyethylene glycol 6000 5.0 grams talc 5.0 grams magnesium stearate 1.8grams demineralized water q.s.

The solid ingredients are first forced through a sieve of 0.6 mm meshwidth. The active ingredient, the lactose, the talc, the magnesiumstearate and half of the starch then are mixed. The other half of thestarch is suspended in 40 milliliters of water and this suspension isadded to a boiling solution of the polyethylene glycol in 100milliliters of water. The resulting paste is added to the pulverulentsubstances and the mixture is granulated, if necessary with the additionof water. The granulate is dried overnight at 35° C., forced through asieve of 1.2 mm mesh width and compressed to form tablets ofapproximately 6 mm diameter which are concave on both sides.

Example 16

Tablets, each containing 100 milligrams of active ingredient, can beprepared in the following manner:

Constituents (for 1000 tablets)

active ingredient 100.0 grams lactose 100.0 grams wheat starch 47.0grams magnesium stearate 3.0 grams

All the solid ingredients are first forced through a sieve of 0.6 mmmesh width. The active ingredient, the lactose, the magnesium stearateand half of the starch then are mixed. The other half of the starch issuspended in 40 milliliters of water and this suspension is added to 100milliliters of boiling water. The resulting paste is added to thepulverulent substances and the mixture is granulated, if necessary withthe addition of water. The granulate is dried overnight at 35° C.,forced through a sieve of 1.2 mm mesh width and compressed to formtablets of approximately 6 mm diameter which are concave on both sides.

EXAMPLE 17

Tablets for chewving, each containing 75 milligrams of activeingredient, can be prepared in the following manner:

Composition (for 1000 tablets).

active ingredient 75.0 grams mannitol 230.0 grams lactose 150.0 gramstalc 21.0 grams glycine 12.5 grams stearic acid 10.0 grams saccharin 1.5grams 5% gelatin solution q.s.

All the solid ingredients are first forced through a sieve of 0.25 mmmesh width. The mannitol and the lactose are mixed, granulated with theaddition of gelatin solution, forced through a sieve of 2 mm mesh width,dried at 50° C. and again forced through a sieve of 1.7 mm mesh width.The active ingredient, the glycine and the saccharin are carefullymixed, the mannitol, the lactose granulate, the stearic acid and thetalc are added and the whole is mixed thoroughly and compressed to formtablets of approximately 10 mm diameter which are concave on both sidesand have a breaking groove on the upper side.

EXAMPLE 18

Tablets, each containing 10 milligrams of active ingredient, can beprepared in the following manner:

Composition (for 1000 tablets)

active ingredient 10.0 grams lactose 328.5 grams corn starch 17.5 gramspolyethylene glycol 6000 5.0 grams talc 25.0 grams magnesium stearate4.0 grams demineralized water q.s.

The solid ingredients are first forced through a sieve of 0.6 mm meshwidth. Then the active ingredient, lactose, talc, magnesium stearate andhalf of the starch are intimately mixed. The other half of the starch issuspended in 65 milliliters of water and this suspension is added to aboiling solution of the polyethylene glycol in 260 milliliters of water.The resulting paste is added to the pulverulent substances, and thewhole is mixed and granulated, if necessary with the addition of water.The granulate is dried overnight at 35° C., forced through a sieve of1.2 mm mesh width and compressed to form tablets of approximately 10 mmdiameter which are concave on both sides and have a breaking notch onthe upper side.

EXAMPLE 19

Gelatin dry-filled capsules, each containing 100 milligrams of activeingredient, can be prepared in the following manner:

Composition (for 1000 capsules)

active ingredient 100.0 grams microcrystalline cellulose 30.0 gramssodium lauryl sulphate 2.0 grams magnesium stearate 8.0 grams

The sodium lauryl sulphate is sieved into the active ingredient througha sieve of 0.2 mm mesh width and the two components are intimately mixedfor 10 minutes. The microcrystalline cellulose is then added through asieve of 0.9 mm mesh width and the whole is again intimately mixed for10 minutes. Finally, the magnesium stearate is added through a sieve of0.8 mm width and, after mixing for a further 3 minutes, the mixture isintroduced in portions of 140 milligrams each into size 0 (elongated)gelatin dry-fill capsules.

EXAMPLE 20

A 0.2% injection or infusion solution can be prepared, for example, inthe following manner: active ingredient 5.0 grams sodium chloride 22.5grams phosphate buffer pH 7.4 300.0 grams demineralized water qs 2500.0milliliters

The active ingredient is dissolved in 1000 milliliters of water andfiltered through a microfilter or slurried in 1000 mL of H₂O. The buffersolution is added and the whole is made up to 2500 milliliters withwater. To prepare dosage unit forms, portions of 1.0 or 2.5 milliliterseach are introduced into glass ampoules (each containing respectively2.0 or 5.0 milligrams of active ingredient).

1. A compound of the formula:

wherein: R⁵ is the divalent residue of pyrrolidine, wherein the twobonds of the divalent residue are on vicinal ring carbon atoms; R⁶ is—CO—, —CH₂—, or —CH₂CO—; R⁷ is (i) cyclic or bicyclic alkyl of 4 to 12carbon atoms; (ii) pyridyl; (iii) phenyl substituted with one or moresubstituents each selected independently of the other from nitro, cyano,trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,carbamoyl, acetoxy, carboxy, hydroxy, amino, straight or branched-alkylof 1 to 10 carbon atoms, straight or branched alkoxy of 1 to 10 carbonatoms, or halo; (iv) benzyl substituted with one to three substituentseach selected independently from the group consisting of nitro, cyano,trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,carbamoyl, acetoxy, carboxy, hydroxy, amino, alkyl of 1 to 4 carbonatoms, alkoxy of 1 to 10 carbon atoms, or halo; (v) naphthyl; or (vi)benzyloxy; Y is —COX, —C≡N, —OR⁸, alkyl of 1 to 5 carbon atoms, or aryl;X is —NH₂, —OH, —NHR, —R⁹, —OR⁹, or alkyl of 1 to 5 carbon atoms; R⁸ ishydrogen or lower alkyl; R⁹ is alkyl or benzyl; and, n has a value of 0,1, 2, or
 3. 2. A compound according to claim 1 wherein Y is —C⇓N or—CO(CH₂)_(m) CH₃ in which m has a value of 0, 1, 2, or 3; and
 3. Acompound of the formula:

in which: one of R¹ and R² is R³—X— and the other is hydrogen, nitro,cyano, trifluoromethyl, carbethoxy, carbomethoxy, carbopropoxy, acetyl,carbamoyl, acetoxy, carboxy, hydroxy, amino, lower alkyl, lower alkoxy,halo, or R³—X—; R³ is monocycloalkyl of up to 10 carbon atoms,polycycloalkyl of up to 10 carbon atoms, or benzocyclic alkyl of up to10 carbon atoms; X is —CH₂ — or —O—; R⁵ is the vicinally divalentresidue of pyrrolidine, wherein the two bonds of the divalent residueare on vicinal ring carbon atoms; R⁶ is —CO—, —CH₂—, or —CH₂CO—; Y is—COX, —C≡N, —OR⁸, alkyl of 1 to 5 carbon atoms, or aryl; X is —NH₂, —OH,—NHR, —R⁹, —OR⁹, or alkyl of 1 to 5 carbon atoms; R⁸ is hydrogen orlower alkyl; R⁹ is alkyl or benzyl; and, n has a value of 0, 1, 2,or3.4. A compound according to claim 3 wherein one of R¹ and R² is R³—O— andthe other is lower alkyl, lower alkoxy, or R³—O—; R³ is cyclic orbicyclic alkyl of up to 10 carbon atoms; R⁵ is the the vicinallydivalent residue of pyrrolidine, wherein the two bonds of the divalentresidue are on vicinal ring carbon atom; R⁶ is —CO— or —CH₂—; Y is —COX;X is —NH₂, —OH, —NHR, —R⁹, or —OR⁹; R⁹ is alkyl or benzyl; and n has avalue of 1 or
 2. 5. A compound according to claim 4 wherein one of R¹and R² is R³—O— and the other is methoxy or ethoxy; R³ is cyclopentyl orcyclohexyl; R⁶ is —CO— or —CH₂—; Y is —COX; X is —NH₂, —OH, or —OR⁹; R⁹is methyl or ethyl; and n has a value of
 1. 6. A compound according toclaim 3 wherein one of R¹ and R² is R³—O— and the other is lower alkyl,lower alkoxy, or R³—O—; R³ is cyclic or bicyclic alkyl of up to 10carbon atoms; R⁵ is the vicinally divalent residue of pyrrolidine; R⁶ is—CO— or —CH₂—; Y is —COX; X is —NH₂, —OH, —NHR, —R⁹, or —OR⁹; R⁹ isalkyl or benzyl; and n has a value of 1 or
 2. 7. A compound according toclaim 4 wherein one of R¹ and R² is R3—O— and the other is methoxy orethoxy; R³ is cyclopentyl or cyclohexyl; R⁶ is —CO— or —CH₂—; Y is —COX;X is —NH₂, —OH, or —OR⁹; R⁹ is methyl or ethyl; and n has a value of 1.8. The method of inhibiting phosphodiesterase in a mammal whichcomprises administering thereto an effective amount of a compound ofclaim
 3. 9. The method of inhibiting TNFα. in a mammal which comprisesadministering thereto an effective amount of a compound of claim
 3. 10.A pharmaceutical composition comprising an amount of a compoundaccording to claim 3 effective upon single or multiple dosage sufficientto inhibit phosphodiesterase or TNFα.
 11. The method of inhibitingphosphodiesterase in a mammal which comprises administering thereto aneffective amount of a compound of claim
 1. 12. The method of inhibitingTNFα. in a mammal which comprises administering thereto an effectiveamount of a compound of claim
 1. 13. A pharmaceutical compositioncomprising an amount of a compound according to claim 1 effective uponsingle or multiple dosage sufficient to inhibit phosphodiesterase orTNFα.